Leptin ELISA Kit (Human) (OKBB00206)

Catalog No: OKBB00206 (Formerly GWB-ZZD030)

Size:96 Tests

Human LEP ELISA Kit standard curve

Product Info

• Predicted Species Reactivity: Human
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's human Leptin ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Leptin has been precoated onto 96-well plates. Standards(E.coli, V22-C167) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Leptin is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The absorbance of yellow is proportional to the human Leptin amount of sample captured in plate.
• ELISA Kit Range: 62.5 pg/ml - 4,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	1026	2535	3542	990	2674	3305
  St. Dev.	37.96	81.12	123.97	57.42	120.33	125.59
  %CV	3.69	3.2	3.5	5.8	4.5	3.8

• ELISA Kit Component:
  • Lyophilized recombinant human IL-12(p40) standard: 10 ng/tube x 2
  • Anti-human IL-12(p40) Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-human IL-12(p40) antibody: 130 uL, dilution 1:100
  • Antibody diluent buffer: 12 mL
  • Avidin-Biotin-Peroxidase Complex (ABC): 130 uL, dilution 1:100
  • ABC diluent buffer: 12 mL
  • TMB color developing agent: 10 mL
  • TMB stop solution: 10 mL
• Additional Information: Range: 62.5pg/ml-4000pg/ml
  Principle: Aviva’s human Leptin ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human Leptin specific-specific monoclonal antibodies were pre-coated onto 96-well plates. The human specific detection monoclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human Leptin amount of sample captured in plate.
  Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot
  experiment using standards and a small number of samples is recommended.
  2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
  3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
  4. Duplicate well assay is recommended for both standard and sample testing.
  5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
  6. Don’t reuse tips and tubes to avoid cross contamination.
  7. To avoid to use the reagents from different batches together.
  8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the
  marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before
  using.
  Background: Leptin (or obese, OB) is a circulating hormone that is expressed abundantly and specifically in the adipose tissue.1 Human ob protein is a 166-amino acid polypeptide with a putative signal sequence and is 84 and 83% homologous to the mouse and rat ob proteins, respectively.2 Northern blot analysis reveals that OB RNA is present at a high level in adipose tissue but at much lower levels in placenta and heart.3 Indeed, Leptin is first described as an adipocyte-derived signalling factor, which, after interaction with its receptors, induces a complex response including control of bodyweight and energy expenditure. Leptin seems in addition to its role in metabolic control to have important roles in reproduction and neuroendocrine signalling.4 Leptin is a powerful inhibitor of bone formation in vivo.5 Comparative mapping of mouse and human DNA indicates that the ob gene is located within a region of mouse chromosome 6 that is homologous to a portion of human chromosome 7q.3 The standard product used in this kit is recombinant human Leptin.
• Reconstitution and Storage: Store at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
• Sample Type: cell culture supernates, serum and plasma (heparin, EDTA)
• Sensitivity: <10 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine
• Reference: 1. Masuzaki, H.; Ogawa, Y.; Sagawa, N.; Hosoda, K.; Matsumoto, T.; Mise, H.; Nishimura, H.; Yoshimasa, Y.; Tanaka,
  I.; Mori, T.; Nakao, K. Nonadipose tissue production of Leptin: Leptin as a novel placenta-derived hormone in
  humans. Nature Med. 3: 1029-1033, 1997.
  2. Masuzaki, H.; Ogawa, Y.; Isse, N.; Satoh, N.; Okazaki, T.; Shigemoto, M.; Mori, K.; Tamura, N.; Hosoda, K.;
  Yoshimasa, Y.; Jingami, H.; Kawada, T.; Nakao, K. Human obese gene expression: adipocyte-specific expression
  and regional differences in the adipose tissue. Diabetes 44: 855-858, 1995.
  3. Green, E. D.; Maffei, M.; Braden, V. V.; Proenca, R.; DeSilva, U.; Zhang, Y.; Chua, S. C., Jr.; Leibel, R. L.;
  Weissenbach, J.; Friedman, J. M. The human obese (OB) gene: RNA expression pattern and mapping on the
  physical, cytogenetic, and genetic maps of chromosome 7. Genome Res. 5: 5-12, 1995.
  4. Auwerx, J.; Staels, B. Leptin. Lancet 351: 737-742, 1998.
  5. Elefteriou, F.; Takeda, S.; Ebihara, K.; Magre, J.; Patano, N.; Ae Kim, C.; Ogawa, Y.; Liu, X.; Ware, S. M.; Craigen,
  W. J.; Robert, J. J.; Vinson, C.; Nakao, K.; Capeau, J.; Karsenty, G. Serum Leptin level is a regulator of bone mass.
  Proc. Nat. Acad. Sci. 101: 3258-3263, 2004.
• Specificity: Natural and recombinant human Leptin
• Immunogen: Expression system for standard: E.coli; Immunogen sequence: V22-C167
• Description: For quantitative detection of human Leptin in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Target Info

• Gene Symbol: LEP
• Gene Full Name: leptin
• Alias Symbols: FLJ94114, LEP, LEP_HUMAN, Leptin, Leptin Murine Obesity Homolog, Leptin Precursor Obesity Factor, OB, Obese protein, Obesity factor, Obesity homolog mouse, Obesity murine homolog leptin, OBS
• NCBI Gene Id: 3952
• Protein Name: Leptin 
• Description of Target: Leptin (or obese, OB) is a circulating hormone that is expressed abundantly and specifically in the adipose tissue.1 Human ob protein is a 166-amino acid polypeptide with a putative signal sequence and is 84 and 83% homologous to the mouse and rat ob proteins, respectively.2 Northern blot analysis reveals that OB RNA is present at a high level in adipose tissue but at much lower levels in placenta and heart.3 Indeed, Leptin is first described as an adipocyte-derived signalling factor, which, after interaction with its receptors, induces a complex response including control of bodyweight and energy expenditure. Leptin seems in addition to its role in metabolic control to have important roles in reproduction and neuroendocrine signalling.4 Leptin is a powerful inhibitor of bone formation in vivo.5 Comparative mapping of mouse and human DNA indicates that the ob gene is located within a region of mouse chromosome 6 that is homologous to a portion of human chromosome 7q.3 The standard product used in this kit is recombinant human Leptin.
• Uniprot ID: P41159
• Protein Accession #: NP_000221.1
• Nucleotide Accession #: NM_000230.2